Cesium Chloride Preparation of DNA: Difference between revisions
added concentration to Acetate Buffer |
updated cesium concentration |
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#Load into two ultracentrifuge tubes then add 25 uL EtBR to each tube | #Load into two ultracentrifuge tubes then add 25 uL EtBR to each tube | ||
#Centrifuge 60 000 RPM (250 000g) O/N in NVT90 rotor | #Centrifuge 60 000 RPM (250 000g) O/N in NVT90 rotor | ||
#Withdraw the upper band with a 18g needle and reload into 1g/ | #Withdraw the upper band with a 18g needle and reload into 0.1g/mL CsCl for second spin | ||
#Remove band and extract EtBr with butanol until clear | #Remove band and extract EtBr with butanol until clear | ||
#Dialyse overnight in 2L of TE | #Dialyse overnight in 2L of TE | ||
#Precipitate with 3 vol absolute ethanol and 1/10 vol of 3M Acetate pH 7 | #Precipitate with 3 vol absolute ethanol and 1/10 vol of 3M Acetate pH 7 | ||
#Resuspend to a final concentration of 5-10 mg/mL in TE | #Resuspend to a final concentration of 5-10 mg/mL in TE | ||